Modelling GATA2 immunodeficiency and the progression towards Myelodysplastic syndrome (MDS) and Acute myeloid leukaemia (AML)

Heterozygous germline mutations in the GATA2 gene gives rise to an immunodeficiency syndrome characterised by cytopenias, severe infections and a predisposition towards haematological maligancies such as myeloidysplastic syndrome (MDS) and acute myeloid leukemia (AML). Clinical data from GATA2 mutated families reveal a variable age at presentation, a variable disease phenotype and a proportion of asymptomatic carriers, suggesting incomplete penetrance. Acquisition of secondary hits such as somatic mutations in the ASXL1 gene and cytogenetic abnormalities such as monosomy 7 and trisomy 8 are commonly found in patients that have progressed towards MDS/AML. Perturbation in the immune system of these patients leads to persistent inflammation. Indeed, scRNA-seq from patient samples showed immune and inflammatory pathway dysregulation at the HSC level, which has also been confirmed in our lab in HSCs from Gata2 heterozygous mice. Whilst there has been much research over the years investigating the loss of GATA2 in a variety of mouse and human models, many have failed to recapitulate key aspects of the disease. Using human iPSCs, primary cord blood CD34+ cells and transgenic mouse models, GATA2 immunodeficiency was modelled along with further investigations into inflammation as a potential driver of disease progression. In this thesis, CRISPR/Cas9 and CRISPR base editors were used to engineer specific GATA2 mutations - T354M and R361C – into human iPSC lines followed by truncating ASXL1 mutations to model GATA2 immunodeficiency and its progression towards myeloid malignancy. Unfortunately, due to technical and time limitations (including COVID19) this work was unable to be taken to fruition. Interestingly, in vitro bacterial stimulation selectively increased CFU output and CD14 expression in GATA2 knockdown cord blood CD34+ cells. In addition, chronic inflammation led to splenomegaly and granuloma formation along with a reduction of Gr1+ Mac1+ cells in the BM of mice with haematopoietic-specific deletion of a single Gata2 allele. Finally, in comparison with steady-state conditions where Gata2- heterozygous HSC levels were reduced by half, HSC numbers remained unchanged while control HSCs were reduced by half upon inflammation, indicating that Gata2 heterozygosity enabled HSCs mechanism to better adapt to inflammation. Taken together, this work highlights that a multi-model approach is desirable to study GATA2 familial MDS/AML and that inflammation is an important mediator for disease progression in GATA2 immunodeficiency syndromes thus opening new avenues for disease modelling and therapeutic intervention

The effect of administration of fenbendazole on the microbial hindgut population of the horse

Anthelmintics are used as anti-worming agents. Although known to affect their target organisms, nothing has been published regarding their effect on other digestive tract organisms, or metabolites produced by them. The current work investigated effects of fenbendazole, a benzimidazole anthelmintic, on bacteria and ciliates in the equine digestive tract, and their major metabolites. Animals receiving anthelmintic treatment, had high faecal egg counts, relative to controls. Analysis was performed over two weeks, with temporal differences detected in bacterial populations, but with no other significant differences detected. This suggests fenbendazole has no detectable effect on organisms other than its targets. Moreover it does not appear to make a contribution to changing the resulting metabolome

Does repetitive task training improve functional activity after stroke? A Cochrane systematic review and meta-analysis.

Repetitive task training resulted in modest improvement across a range of lower limb outcome measures, but not upper limb outcome measures. Training may be sufficient to have a small impact on activities of daily living. Interventions involving elements of repetition and task training are diverse and difficult to classify: the results presented are specific to trials where both elements are clearly present in the intervention, without major confounding by other potential mechanisms of action

Exploring women's priorities for the potential consequences of a gestational diabetes diagnosis: a pilot community jury

Background: There is no international diagnostic agreement for gestational diabetes mellitus (GDM). In 2014, Australia adopted a new definition and testing procedure. Since then, significantly more women have been diagnosed with GDM but with little difference in health outcomes. We explored the priorities and preferences of women potentially impacted by a GDM diagnosis. Method: We recruited 15 women from the Gold Coast, Australia, to participate in a pilot community jury (CJ). Over two days, the women deliberated on the following: (a) which important consequences of a diagnosis of GDM should be considered when defining GDM?; (b) what should Australian health practitioners call the condition known as GDM? Results: Eight women attended the pilot CJ, and their recommendations were a consensus. Women were surprised that the level of risk for physical harms was low but emotional harms were high. The final ranking of important consequences (high to low) was as follows: women's negative emotions; management burden of GDM; overmedicalized pregnancy; minimizing infant risks; improving lifestyle; and macrosomia. To describe the four different clinical states of GDM, the women chose three different labels. One was GDM. Conclusions: The women from this pilot CJ prioritized the consequences of a diagnosis of GDM differently from clinicians. The current glucose threshold for GDM in Australia is set at a cut-point for adverse risks including macrosomia and neonatal hyperinsulinaemia. Definitions and guideline panels often fail to ask the affected public about their values and preferences. Community voices impacted by health policies should be embedded in the decision-making process

Beyond the MHC: A canine model of dermatomyositis shows a complex pattern of genetic risk involving novel loci

Juvenile dermatomyositis (JDM) is a chronic inflammatory myopathy and vasculopathy driven by genetic and environmental influences. Here, we investigated the genetic underpinnings of an analogous, spontaneous disease of dogs also termed dermatomyositis (DMS). As in JDM, we observed a significant association with a haplotype of the major histocompatibility complex (MHC) (DLA-DRB1*002:01/-DQA1*009:01/-DQB1*001:01), particularly in homozygosity (P-val = 0.0001). However, the high incidence of the haplotype among healthy dogs indicated that additional genetic risk factors are likely involved in disease progression. We conducted genome-wide association studies in two modern breeds having common ancestry and detected strong associations with novel loci on canine chromosomes 10 (P-val = 2.3X10-12) and 31 (P-val = 3.95X10-8). Through whole genome resequencing, we identified primary candidate polymorphisms in conserved regions of PAN2 (encoding p.Arg492Cys) and MAP3K7CL(c.383_392ACTCCACAAA\u3eGACT) on chromosomes 10 and 31, respectively. Analyses of these polymorphisms and the MHC haplotypes revealed that nine of 27 genotypic combinations confer high or moderate probability of disease and explain 93% of cases studied. The pattern of disease risk across PAN2 and MAP3K7CL genotypes provided clear evidence for a significant epistatic foundation for this disease, a risk further impacted by MHC haplotypes. We also observed a genotype-phenotype correlation wherein an earlier age of onset is correlated with an increased number of risk alleles at PAN2 and MAP3K7CL. High frequencies of multiple genetic risk factors are unique to affected breeds and likely arose coincident with artificial selection for desirable phenotypes. Described herein is the first three-locus association with a complex canine disease and two novel loci that provide targets for exploration in JDM and related immunological dysfunction

‘Single‐handed care’ initiatives and reviews of double‐handed homecare packages: A survey of practices in English local authorities with adult social care responsibilities

International health and social care systems are experiencing unprecedented pressure and demand. ‘Single‐handed care’ initiatives seek to identify whether all or part of a homecare package involving more than one care worker can be safely reduced to a single worker. Little is known about these initiatives across local authorities. The aim of this study was to identify, describe and explain current processes and practices for single‐handed care initiatives and double‐handed homecare reviews. An electronic survey link was sent to each local authority with social care responsibilities in England. The questions covered a range of areas in relation to single‐handed care processes and included a combination of pre‐coded and free‐text responses. Responses were received from 76 (50%) local authorities. Findings were that over 12,000 reviews were reported within a year with a median of 141 (IQR 45–280) from 53 authorities that provided figures. Reviews were usually led by a local authority occupational therapist. On average, 540 min was spent per review, including conducting and organising the review, documentation, and travel. In nearly half the authorities, double handed care remained at least partially in place following at least 80% of the reviews and remained wholly in place following at least 60%. Local authorities also reported some resistance from homecare providers when implementing single‐handed care. The findings have confirmed anecdotal evidence that reviews of double‐handed homecare packages are common practice within local authorities. Given the amount of time taken with these reviews, and paucity of evidence on outcomes for people receiving them, further research should evaluate this

The fables of pity: Rousseau, Mandeville and the animal-fable

Copyright @ 2012 Edinburgh University PressPrompted by Derrida’s work on the animal-fable in eighteenth-century debates about political power, this article examines the role played by the fiction of the animal in thinking of pity as either a natural virtue (in Rousseau’s Second Discourse) or as a natural passion (in Mandeville’s The Fable of the Bees). The war of fables between Rousseau and Mandeville – and their hostile reception by Samuel Johnson and Adam Smith – reinforce that the animal-fable illustrates not so much the proper of man as the possibilities and limitations of a moral philosophy that is unable to address the political realities of the state